Clinical chorioamnionitis at term X: microbiology, clinical signs, placental pathology, and neonatal bacteremia - implications for clinical care.

OBJECTIVES: Clinical chorioamnionitis at term is considered the most common infection-related diagnosis in labor and delivery units worldwide. The syndrome affects 5-12% of all term pregnancies and is a leading cause of maternal morbidity and mortality as well as neonatal death and sepsis. The objectives of this study were to determine the (1) amniotic fluid microbiology using cultivation and molecular microbiologic techniques; (2) diagnostic accuracy of the clinical criteria used to identify patients with intra-amniotic infection; (3) relationship between acute inflammatory lesions of the placenta (maternal and fetal inflammatory responses) and amniotic fluid microbiology and inflammatory markers; and (4) frequency of neonatal bacteremia. METHODS: This retrospective cross-sectional study included 43 women with the diagnosis of clinical chorioamnionitis at term. The presence of microorganisms in the amniotic cavity was determined through the analysis of amniotic fluid samples by cultivation for aerobes, anaerobes, and genital mycoplasmas. A broad-range polymerase chain reaction coupled with electrospray ionization mass spectrometry was also used to detect bacteria, select viruses, and fungi. Intra-amniotic inflammation was defined as an elevated amniotic fluid interleukin-6 (IL-6) concentration ≥2.6 ng/mL. RESULTS: (1) Intra-amniotic infection (defined as the combination of microorganisms detected in amniotic fluid and an elevated IL-6 concentration) was present in 63% (27/43) of cases; (2) the most common microorganisms found in the amniotic fluid samples were Ureaplasma species, followed by Gardnerella vaginalis; (3) sterile intra-amniotic inflammation (elevated IL-6 in amniotic fluid but without detectable microorganisms) was present in 5% (2/43) of cases; (4) 26% of patients with the diagnosis of clinical chorioamnionitis had no evidence of intra-amniotic infection or intra-amniotic inflammation; (5) intra-amniotic infection was more common when the membranes were ruptured than when they were intact (78% [21/27] vs. 38% [6/16]; p=0.01); (6) the traditional criteria for the diagnosis of clinical chorioamnionitis had poor diagnostic performance in identifying proven intra-amniotic infection (overall accuracy, 40-58%); (7) neonatal bacteremia was diagnosed in 4.9% (2/41) of cases; and (8) a fetal inflammatory response defined as the presence of severe acute funisitis was observed in 33% (9/27) of cases. CONCLUSIONS: Clinical chorioamnionitis at term, a syndrome that can result from intra-amniotic infection, was diagnosed in approximately 63% of cases and sterile intra-amniotic inflammation in 5% of cases. However, a substantial number of patients had no evidence of intra-amniotic infection or intra-amniotic inflammation. Evidence of the fetal inflammatory response syndrome was frequently present, but microorganisms were detected in only 4.9% of cases based on cultures of aerobic and anaerobic bacteria in neonatal blood.

Rare case of osteomyelitis caused by Gardnerella vaginalis and Streptococcus parasanguinis in a postmenopausal woman.

Vertebral osteomyelitis is an infection of the vertebrae that can lead to spinal degeneration, most commonly caused by Staphylococcus aureus Here, we report an unusual case of pyogenic osteomyelitis caused by Gardnerella vaginalis and Streptococcus parasanguinis in a 61-year-old postmenopausal woman. The patient presented with a 2-week history of worsening lower back pain and fever and a recent episode of cystitis following re-engagement of sexual activity. Imaging revealed a deterioration of vertebrae discs and spinal canal stenosis at the L3-L4 levels with a formation of abscess in the right psoas muscle. Needle aspiration of the abscess identified G. vaginalis and S. parasanguinis and the patient was successfully treated with a 6-week course of ceftriaxone and metronidazole. This case describes an unusual coinfection of two pathogens that normally reside in the urogenital tract and oral cavity, respectively, and highlights the risk posed when these organisms breach the body's normal barriers.

Cervical Gardnerella vaginalis in women with preterm prelabor rupture of membranes.

OBJECTIVE: To determine the association between microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI) and the cervical prevalence of Gardnerella vaginalis DNA in pregnancies with preterm prelabor rupture of membrane (PPROM). METHOD: In total, 405 women with singleton pregnancies complicated with PPROM were included. Cervical fluid and amniotic fluid samples were collected at the time of admission. Bacterial and G. vaginalis DNA were assessed in the cervical fluid samples using quantitative PCR technique. Concentrations of interleukin-6 and MIAC were evaluated in the amniotic fluid samples. Loads of G. vaginalis DNA ≥ 1% of the total cervical bacterial DNA were used to define the cervical prevalence of G. vaginalis as abundant. Based on the MIAC and IAI, women were categorized into four groups: with intra-amniotic infection (both MIAC and IAI), with sterile IAI (IAI without MIAC), with MIAC without IAI, and without either MIAC or IAI. RESULTS: The presence of the abundant cervical G. vaginalis was related to MIAC (with: 65% vs. without: 44%; p = 0.0004) but not IAI (with: 52% vs. without: 48%; p = 0.70). Women with MIAC without IAI had the highest load of the cervical G. vaginalis DNA (median 2.0 × 104 copies DNA/mL) and the highest presence of abundant cervical G. vaginalis (73%). CONCLUSIONS: In women with PPROM, the presence of cervical G. vaginalis was associated with MIAC, mainly without the concurrent presence of IAI.

Associação entre Vaginose Bacteriana e Anormalidades Citológicas nos Exames Citopatológicos Analisados em um Laboratório Escola de Goiânia-GO / Association Between Bacterial Vaginosis and Cytological Abnormalities in Cytopathological Exams Analyzed in a Laboratory School of Goiânia-GO / Asociación entre Vaginosis Bacteriana y Anormalidades Citológicas en Exámenes Citopatológicos Analizados en una Escuela de Laboratorio de Goiânia-GO

Introdução: A Gardnerella vaginalis facilita a infecção pelo papilomavírus humano (HPV). Objetivo: Verificar a associação entre anormalidades citológicas e presença de Gardnerella vaginalis nos esfregaços cervicovaginais encaminhados ao Laboratório Clínico da Pontifícia Universidade Católica de Goiás (LAC/PUC Goiás) estratificadas por faixa etária. Método: Estudo transversal realizado no LAC/PUC Goiás entre janeiro de 2013 a dezembro de 2015. Para análises estatísticas, a variável idade foi categorizada em ≤39 anos e >40 anos, utilizando o programa IBM SPSS Statistics (Version 2.0, 2011®) para o teste de qui-quadrado (X²), com intervalo de confiança de 95% e valor p<0,05. Resultados: Foram analisados 4.558 exames citopatológicos, a maioria com presença de Lactobacillus spp. (46,97%). A prevalência dos agentes patogênicos foi a Gardnerella vaginalis (79,6%), seguida de Candida spp. (16,8%), Trichomonas vaginalis (2,2%), Herpes simplex (0,4 %) e Chlamydia trachomatis (0,1%). As anormalidades citológicas foram observadas em 9,1%, sendo atypical squamous cells of undetermined significance (ASC-US) 2,57%, low grade squamous intraepithelial lesion (LSIL) 1,78%, atypical squamous cells of undetermined significance cannot exclude high grade squamous intraepithelial lesion (ASC-H) 3,52%, high grade squamous intraepithelial lesion (HSIL) 1,08%, atypical endocervical cells, favor neoplastic (AGC-NEO) 0,22% e carcinoma 0,02%. Houve uma associação significante entre anormalidades citológicas graves e mulheres ≥40 anos, OR 3,01 (IC 95% 2,0-4,58) (p<0,0001). Mulheres ≤40 anos mostraram significância à presença de Gardnerella vaginalis (p<0,0004). Conclusão: Uma elevada prevalência de Gardnerella vaginalis foi encontrada associada com as anormalidades citológicas, principalmente em mulheres sexualmente ativas.

Introduction:Gardnerella vaginalis facilitates human papillomavirus (HPV) infection. Objective: To verify the association between cytological abnormalities and the presence of Gardnerella vaginalis in cervicovaginal smears sent to the Clinical Laboratory of the Pontifical Catholic University of Goiás (LAC/PUC Goiás) stratified by age range. Method: Cross-sectional study carried out at LAC/PUC Goiás from January 2013 to December 2015. For statistical analysis, the variable age was categorized as ≤39 years and >40 years, using the IBM SPSS Statistics program (Version 2.0, 2011®) for the chi-square test (X²), with a 95% confidence interval and p<0.05. Results:4,558 cytopathological exams were analyzed, most of them with the presence of Lactobacillus spp (46.97%). The prevalence of pathogens was Gardnerella vaginalis (79.6%), followed by Candida spp. (16.8%), Trichomonas vaginalis (2.2%), Herpes simplex (0.4%) and Chlamydia trachomatis (0.1%). Cytological abnormalities were observed in 9.1%, being atypical squamous cells of undetermined significance (ASC-US) 2.57%, low grade squamous intraepithelial lesion (LSIL) 1.78%, atypical squamous cells of undetermined significance cannot exclude high intraepithelial lesion (ASC-H) 3.52%, high grade squamous intraepithelial lesion (HSIL) 1.08%, atypical endocervical cells, neoplastic favor (AGC-NEO) 0.22% and carcinoma 0.02%. There was a significant association between severe cytological abnormalities and women >40 years old OR 3.01 (95% CI 2.0-4.58) (p<0.0001). Women ≤40 years old showed the presence of Gardnerella vaginalis (p<0.0004). Conclusion:A high prevalence of Gardnerella vaginalis was found and its association with cytological abnormalities, especially in sexually active women.

Introducción:Gardnerella vaginalis facilita la infección por el virus del papiloma humano (VPH). Objetivo: Verificar la asociación entre anormalidades citológicas y la presencia de Gardnerella vaginalis en frotis cervicovaginales enviadas al Laboratorio Clínico de la Pontificia Universidad Católica de Goiás (LAC/PUC Goiás) estratificadas por grupo de edad. Método: Estudio transversal realizado en LAC/PUC Goiás desde enero de 2013 hasta diciembre de 2015. Para el análisis estadístico, la edad variable se clasificó como ≤39 años y >40 años, utilizando el programa IBM SPSS Statistics (Versión 2.0, 2011®) para la prueba de chi-cuadrado (X²), con un intervalo de confianza del 95% y p <0,05. Resultados: Se analizaron 4.558 exámenes citopatológicos. La prevalencia de Lactobacillusspp. con 46,97%. Los patógenos como Gardnerella vaginalis fueron 79,6%, Candidaspp. 16,8%, Trichomonas vaginalis 2,2%, Herpes simplex 0,4%, y Chlamydia trachomatis 0,1%. Se observaron anormalidades citológicas en 9,1%, con células escamosas atípicas de significado indeterminado (ASC-US) 2,57%, lesión intraepitelial escamosa de bajo grado (LSIL) 1,78%, células escamosas atípicas de significación indeterminada no pueden excluir lesión intraepitelial (ASC-H) 3,52%, lesión intraepitelial escamosa de alto grado (HSIL) 1,08%, células endocervicales atípicas, favor neoplásico (AGC-NEO) 0,22% y carcinoma 0,02%. Hubo una asociación significativa entre anormalidades citológicas severas y mujeres >40 años OR 3,01 (IC 95% 2,0-4,58) (p<0,0001). Las mujeres ≤40 años mostraron la presencia de Gardnerella vaginalis (p<0,0004). Conclusión: Se encontró una alta prevalencia de Gardnerella vaginalis y su asociación con anomalías citológicas, especialmente en mujeres sexualmente activas.

Prevalence and factors associated with Trichomonas vaginalis infection in indigenous Brazilian women.

There is a scarcity of studies on the prevalence of Trichomonas vaginalis (TV) in indigenous populations of Brazil. We conducted a cross-sectional study between January and December 2018, on indigenous women living nearby an urban center of the Midwest region of Brazil and determined the prevalence of TV. Factors associated with TV infection and a comparison of molecular and direct microscopy diagnoses were determined. 241 indigenous women aged above 18 years participated in the study. Cervical and vaginal brush samples were collected to diagnose TV through polymerase chain reaction (PCR). Direct microscopy for detection of TV, and cellular changes was performed. A sociodemographic and behavioral questionnaire was applied at the beginning of the study. All the data were analyzed using Statistical Package for the Social Sciences. The result obtained showed that 27.8% [95% CI: 22.2-33.9] were positive for TV on PCR, while 7.41% [95% CI: 4.1-11] showed positive on direct microscopy. Direct microcopy also found 21 (8.71%) and 8 (3.31%) women infected with Gardnerella vaginalis and Candida albicans, respectively. In addition, 10 women presented atypical squamous cells of unknown significance and 14 lesions suggestive of HPV. Single women, under the age of 30 and who do not use condoms, were found to have a greater chance of getting TV infection. The high prevalence TV found in this population is comparable to highly vulnerable populations, as prisoners, sex workers and women in regions with low socioeconomic levels, moreover, seems to be an underdiagnosis of this infection. Therefore, a routine test program, as well as a review of the diagnostic method used, is encouraged for proper management.

Molecular epidemiology and socio-demographic risk factors of sexually transmitted infections among women in Lebanon.

BACKGROUND: Sexually transmitted infections (STIs) cause a major public health problem that affect both men and women in developing and developed countries. The aim of the study was to estimate the prevalence of 11 STIs among women who voluntarily participated in the study, while seeking gynecological checkup. The existence of an association between the presence of pathogens and symptoms and various sociodemographic risk factors was assessed. METHODS: A total of 505 vaginal and cervical specimens were collected from women above 18 years of age, with or without symptoms related to gynecological infections. Nucleic acid was extracted and samples were tested by real-time PCR for the following pathogens: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Ureaplasma urealyticum, Urealplasma parvum, Trichomonas vaginalis, Mycoplasma hominis, Mycoplasma girerdii, Gardnerella vaginalis, Candida albicans and Human Papillomavirus (HPV). Positive HPV samples underwent genotyping using a microarray system. RESULTS: Of the 505 samples, 312 (62%) were screened positive for at least one pathogen. Of these, 36% were positive for Gardnerella vaginalis, 35% for Ureaplasma parvum, 8% for Candida albicans, 6.7% for HPV, 4.6% for Ureaplasma urealyticum, 3.6% for Mycoplasma hominis, 2% for Trichomonas vaginalis, 0.8% for Chlamydia trachomatis, 0.4% for Mycoplasma girerdii, 0.2% for Mycoplasma genitalium and 0.2% for Neisseria gonorrhoeae. Lack of symptoms was reported in 187 women (37%), among whom 61% were infected. Thirty-four samples were HPV positive, with 17 high risk HPV genotypes (HR-HPV); the highest rates being recorded for types 16 (38%), 18 (21%) and 51 (18%). Out of the 34 HPV positives, 29 participants had HR-HPV. Association with various risk factors were reported. CONCLUSIONS: This is the first study that presents data about the presence of STIs among women in Lebanon and the MENA region by simultaneous detection of 11 pathogens. In the absence of systematic STI surveillance in Lebanon, concurrent screening for HPV and PAP smear is warranted.

Production and characterization of a monoclonal antibody against the sialidase of Gardnerella vaginalis using a synthetic peptide in a MAP8 format.

Bacterial vaginosis is one of the most frequent vaginal infections. Its main etiological agent is Gardnerella vaginalis, which produces several virulence factors involved in vaginal infection and colonization, in particular, sialidase (SLD), a potential clinical biomarker that participates in immune response modulation and mucus degradation. The main objective of this work was the production and evaluation of a monoclonal antibody against G. vaginalis sialidase and its validation in immunoassays. For immunization of mice, a synthetic multiantigenic peptide was used, and hybridomas were generated. After fusion, hybridomas were evaluated for antibody production and cloned by limited dilution. One clone producing IgG1 was selected and characterized by indirect ELISA, dot blot, and Western blot, and we also tested clinical isolates and HeLa cells infected with G. vaginalis. The results showed that the anti-SLD antibody recognized a single protein of ~90 kDa that correlated with the estimated molecular weight of SLD. In addition, anti-SLD antibody recognized SLD from complete bacteria and from culture supernatants of infected Hela cells. In conclusion, our results showed that the anti-SLD antibody recognized SLD from different sources and could be considered a new tool for the diagnosis of bacterial vaginosis. KEY POINTS: • Anti-sialidase mAb was generated using a synthetic peptide • The mAb recognizes synthetic peptide and intact protein from multiple sources • The antibody was characterized by several immunological methods.

The vaginal microbiota associates with the regression of untreated cervical intraepithelial neoplasia 2 lesions.

Emerging evidence suggests associations between the vaginal microbiota (VMB) composition, human papillomavirus (HPV) infection, and cervical intraepithelial neoplasia (CIN); however, causal inference remains uncertain. Here, we use bacterial DNA sequencing from serially collected vaginal samples from a cohort of 87 adolescent and young women aged 16-26 years with histologically confirmed, untreated CIN2 lesions to determine whether VMB composition affects rates of regression over 24 months. We show that women with a Lactobacillus-dominant microbiome at baseline are more likely to have regressive disease at 12 months. Lactobacillus spp. depletion and presence of specific anaerobic taxa including Megasphaera, Prevotella timonensis and Gardnerella vaginalis are associated with CIN2 persistence and slower regression. These findings suggest that VMB composition may be a future useful biomarker in predicting disease outcome and tailoring surveillance, whilst it may offer rational targets for the development of new prevention and treatment strategies.

Apoptosis of vaginal epithelial cells in clinical samples from women with diagnosed bacterial vaginosis.

Bacterial vaginosis (BV) is one of the most common vaginal infections among women of childbearing age. Gardnerella vaginalis (G. vaginalis) is a keystone microorganism present in more than 95% of all BV cases. The first step of the infection process in BV is mediated by interaction of microorganisms with epithelial cells (ECs). However, the role of these cells in BV pathogenesis is largely unknown. The present study aimed to investigate the vaginal EC response during BV. Twenty healthy women and 34 women with BV were enrolled in this study. The number of ECs in the vaginal swab was counted and analyzed for intracellular signals and apoptosis by flow cytometry. Cell damage was evaluated by lactate dehydrogenase assay. Compared to that in healthy donors, the percentage of exfoliated vaginal ECs was increased in women with BV, and an absence of neutrophils was observed in both groups. Activation signals, such as p-IκBα and c-Fos were unmodulated in the vaginal ECs of women with BV. Moreover, EC damage and apoptosis were significantly increased in patients with BV. Apoptosis was related to caspase-3 activation and the presence of G. vaginalis. This study provides the first evidence of a direct involvement of G. vaginalis in the apoptotic process of vaginal ECs during BV. This effect was mediated by caspase-3 activation, and G. vaginalis appeared to be one of causes for inducing EC apoptosis in BV. Hence, our findings suggest a possible explanation for the increased exfoliation of ECs in the vagina during BV.

Cervical cancer and vaginal microbiota changes.

Microbial changes in vaginal ecosystem may accelerate the process of cervical carcinogenesis. The developed cervical cancer can lead to changes in the vaginal microbiota. The aim of our study is to determine the vaginal microbiota changes at women with FIGO I stage cervical cancer. We conducted an open, single-site survey in the Department of Gynecology of the Military Medical Academy in Sofia, Bulgaria, from 2014 to 2019 year. The study included a total of 32 women aged 38-55 years with clinical and pathology established cervical cancer (FIGO I stage). The underlying vaginal DNA microbiological test indicated presence or absence of bacterial vaginosis, other vaginal infections or normal vaginal microbiota. Of 32 (100%) women enrolled in our study, 19 (59.4%) was with FIGO IA stage cervical cancer and 13 (40.6%) with IB stage. Disturbances of vaginal microbiota in we found at 23 (71.9%) of women with cervical cancer included in our study. At the rest of 9 (28.1%) women we found out normal vaginal microbiota. Bacterial vaginosis was determined clinically and microbiologically in 15 (46.9%) women enrolled in the study. Aerobic vaginitis caused by Streptococcus species we establish at 4 (12.5%) of women. Trichomonas vaginalis infection have 1 (3.1%) women and Candida Albicans the last one 1 (3.1%) from this group with disturbed vaginal microbial balance. Bacterial dysbacteriosis, characterized by a predominance of Gardnerella vaginalis alone or in complex with other anaerobic bacteria, aerobic vaginitis and other sexually transmitted vaginal pathogens from one side and a concomitant paucity of vaginal Lactobacillus species may be an HPV-dependent cofactor for cervical neoplasia development. Only with this single observation it is difficult to confirm that vaginal microbiota dysbiosis contributes to HPV infection and carcinogenesis.

Vaginosis-associated bacteria and its association with HPV infection. / Bacterias relacionadas con vaginosis bacteriana y su asociación a la infección por virus del papiloma humano.

BACKGROUND AND OBJECTIVE: Cervical cancer is an important health problem in our country. It is known that there are several risk factors for this neoplasm, and it has been suggested that cervical microbiome alterations could play a role in the development and progress of cancer. Bacterial vaginosis associated bacteria such as Atopobium vaginae and Gardnerella vaginalis has been suggested as potential risk factor for cervical lesions and cervical cancer. MATERIAL AND METHODS: DNA from 177 cervical scraping samples was studied: 104 belonged to women without cytological or colposcopic alterations and 73 samples from precursor lesions with previous human papillomavirus (HPV) infection history. All samples were screened for Atopobium vaginae, Gardnerella vaginalis and HPV by PCR. RESULTS: High HPV prevalence was found in precursor samples, and 30% of samples without lesions were positive for HPV. Virtually all samples contained sequences of both bacteria, and interestingly, there was not HPV association observed; these results could suggest that these microorganisms could be part of the cervical microbiome in Mexican population. CONCLUSIONS: The results obtained indicate that the bacteria analysed could be part of normal biome in Mexican women, suggesting a potential reconsideration of the pathogen role of these microorganisms.

Co-infection of sexually transmitted pathogens and Human Papillomavirus in cervical samples of women of Brazil.

BACKGROUND: Some sexually transmitted infectious agents, such as Chlamydia trachomatis and Herpes simplex, cause local inflammation, and could contribute to Human Papillomavirus (HPV) and cervical lesion progression. Thus, the aim of this study was to determine any association between the presence of microorganisms of gynecological importance, sexual behavior, clinical and demographical variables to the development and progress of cervical lesions. METHODS: One hundred and thirty-two women between 14 and 78 years and living at Vitória da Conquista, Bahia, Brazil, were included (62 individuals with cervical lesions and 70 without lesions). They answered a questionnaire to provide data for a socioeconomic and sexual activity profile. Samples of cervical swabs were collected and analyzed by PCR to detect genital microorganisms and HPV. Quantitative PCR was used to detect and quantify Ureaplasma urealyticum and Ureaplasma parvum. Univariate and multiple logistic regression were performed to measure the association with the cervical lesions, and an odds ratio (OR) with 95% confidence intervals (95%CI) were calculated. The Mann-Whitney U test was also used to compare the microorganism load in the case and control groups. The significance level was 5% in all hypotheses tested. RESULTS: Cervical lesions were associated with: women in a stable sexual relationship (OR = 14.21, 95%CI = 3.67-55.018), positive PCR for HPV (OR = 16.81, 95%CI = 4.19-67.42), Trichomonas vaginalis (OR = 8.566, 95%CI = 2.04-35.94) and Gardnerella vaginalis (OR = 6.13, 95%CI = 1.53-24.61), adjusted by age and qPCR for U. parvum. U. parvum load showed a statistical difference between the case and control groups (p-value = 0.002). CONCLUSION: Variables such as stable relationship, HPV, T. vaginalis, G. vaginalis were associated with cervical lesions in epidemiological studies. U. parvum load was higher in woman with cervical lesions compared with women without lesions. Additional studies are needed to better understand the role of these factors in cervical lesion development.

Association between statin use, the vaginal microbiome, and Gardnerella vaginalis vaginolysin-mediated cytotoxicity.

BACKGROUND: Bacterial vaginosis (BV) is the leading dysbiosis of the vaginal microbiome. The pathways leading towards the development of BV are not well understood. Gardnerella vaginalis is frequently associated with BV. G. vaginalis produces the cholesterol-dependent cytolysin (CDC), vaginolysin, which can lyse a variety of human cells and is thought to play a role in pathogenesis. Because membrane cholesterol is required for vaginolysin to function, and because HMG-CoA reductase inhibitors (statins) affect not only serum levels of cholesterol but membrane levels as well, we hypothesized that statins might affect the vaginal microbiome. METHODS: To investigate the relationship between use of the statins and the vaginal microbiome, we analyzed 16S rRNA gene taxonomic surveys performed on vaginal samples from 133 women who participated in the Vaginal Human Microbiome Project and who were taking statins at the time of sampling, 152 women who reported high cholesterol levels but were not taking statins, and 316 women who did not report high cholesterol. To examine the effect of statins on the cytolytic effect of vaginolysin, the cholesterol-dependent cytolysin (CDC) produced by Gardnerella vaginalis, we assessed the effect of simvastatin pretreatment of VK2E6/E7 vaginal epithelial cells on vaginolysin-mediated cytotoxicity. RESULTS: The mean proportion of G. vaginalis among women taking statins was significantly lower relative to women not using statins. Women using statins had higher mean proportions of Lactobacillus crispatus relative to women with normal cholesterol levels, and higher levels of Lactobacillus jensenii relative to women with high cholesterol but not taking statins. In vitro, vaginal epithelial cells pretreated with simvastatin were relatively resistant to vaginolysin and this effect was inhibited by cholesterol. CONCLUSIONS: In this cross-sectional study, statin use was associated with reduced proportions of G. vaginalis and greater proportions of beneficial lactobacilli within the vaginal microbiome. The negative association between statin use and G. vaginalis may be related to inhibition of vaginolysin function.

ENDOMETRIAL POLYPS IN WOMEN OF REPRODUCTIVE AGE: CLINICAL AND PATHOGENE-TIC VARIATIONS.

The aim of the study was to study the relationship between the morphofunctional characteristics of the endometrium, hormonal homeostasis and microbiocenosis of the reproductive system in patients with endometrial polyps. The study involved 130 patients aged 18-35 years: 34 patients with endometrial polyps, 30 patients with micropolyps, 36 patients with endometrial polyps and micropolyps, 30 healthy women of the control group. Hysteroscopy was performed for women who had been suspected for endometrial polyps and who had infertility or repeated recurrent miscarriages. Endometrial samples from healthy women were obtained by aspiration biopsy. The endometrial sections were immunostained with monoclonal antibodies against the specific markers of plasmacytes (CD138), NK cells (CD56, CD16), pan-leukocytes (CD45), macrophages (CD68), cellular marker for proliferation (Ki-67), ER, PR. Bacteriological examination of the endometrium was performed by PCR and by cultivating aerobic and anaerobic microorganisms on special growth media. In all groups of women the content in blood serum for 3-5 day of a menstrual cycle of gonadotropic hormones (FSH, LH) and sex steroid hormones (estradiol, prolactin) was studied, for 21 days of a cycle estimated the content of progesterone. Level of an expression of receptors of progesterone and estrogen estimated in endometrium and at EP, also in І a cycle phase. Highlighted are separate clinical and pathogenetic variations of endometrial polyps: isolated polyps, micropolyps, polyps in conjunction with micropolyps. In the course of study, it was found that progesterone deficiency and local immune imbalance with severe hypofunctional NK cells against viral and fungal infestations result in excessive endometrial cell proliferation and development of an isolated polyp. The case of a polyp merging with micropolyps potentiates an active inflammatory process alongside all of the mechanisms mentioned above. Micropolyps as a macroscopic manifestation of an active inflammatory process in chronic endometritis are characterized by focal infiltrates of leukocytes (CD45), macrophages (CD68), plasmacells (CD138) and NK (CD56) cells, whose activity leads to excess abnormal proliferation of endometrium, even in the absence of hormone receptor disorders.

Genital Injury Signatures and Microbiome Alterations Associated With Depot Medroxyprogesterone Acetate Usage and Intravaginal Drying Practices.

Background: Increasing evidence suggests depot medroxyprogesterone acetate (DMPA) and intravaginal practices may be associated with human immunodeficiency virus (HIV-1) infection risk; however, the mechanisms are not fully understood. This study evaluated the effect of DMPA and intravaginal practices on the genital proteome and microbiome to gain mechanistic insights. Methods: Cervicovaginal secretions from 86 Kenyan women, including self-reported DMPA users (n = 23), nonhormonal contraceptive users (n = 63), and women who practice vaginal drying (n = 46), were analyzed using tandem-mass spectrometry. Results: We identified 473 human and 486 bacterial proteins from 18 different genera. Depot medroxyprogesterone acetate use associated with increased hemoglobin and immune activation (HBD, HBB, IL36G), and decreased epithelial repair proteins (TFF3, F11R). Vaginal drying associated with increased hemoglobin and decreased phagocytosis factors (AZU1, MYH9, PLAUR). Injury signatures were exacerbated in DMPA users who also practiced vaginal drying. More diverse (H index: 0.71 vs 0.45; P = .009) bacterial communities containing Gardnerella vaginalis associated with vaginal drying, whereas DMPA showed no significant association with community composition or diversity. Conclusions: These findings provide new insights into the impact of DMPA and vaginal drying on mucosal barriers. Future investigations are needed to confirm their relationship with HIV risk in women.

Frequency of detection of Gardnerella vaginalis in vaginal smears in the Upper Carniola region.

INTRODUCTION: Bacterial vaginosis is of clinical interest because of its possible causal relationship with complications during pregnancy, postpartum, and complications after surgery. METHODS: Gram stain for clue cells and Gardnerella vaginalis culture methods were evaluated retrospectively in a microbiological medical laboratory for the first half of 2015. We were interested in the proportion of G. vaginalis bacteria isolated from genital samples, correlation with Gram-staining presence of clue cells, referral clinical diagnosis, and pregnancy. RESULTS: In the first half of 2015 we received 358 vaginal specimens; 82% of them had a referral clinical diagnosis of colpitis, cervicitis, or vaginal discharge; 40% were pregnant women. G. vaginalis was isolated from 14% of vaginal specimens, and 52% of these came from pregnant patients. Gram stain clue cells and isolation of G. vaginalis matched in 86%. CONCLUSIONS: For diagnosing bacterial vaginosis in clinical practice, standard clinical criteria, Gram staining of vaginal discharge smear, and/or isolation of G. vaginalis are used. Isolation of G. vaginalis without clue cells is reported only in cases in which bacterial growth is predominant. The results of our studies confirm that isolating G. vaginalis helps confirm the diagnosis of bacterial vaginosis.

Combined effect of diet and cervical microbiome on the risk of cervical intraepithelial neoplasia.

BACKGROUND & AIMS: Several food groups or dietary factors and the cervical microbiota may be involved in cervical carcinogenesis, but the evidence is not clear yet. We aimed to assess the association between dietary pattern and cervical intraepithelial neoplasia (CIN) and the combined effect of dietary pattern and cervical microbiome on the risk of CIN. METHODS: The cervical microbiota and diet assessed by pyrosequencing and a food-frequency questionnaire, respectively, of 65 women with CIN and 72 control women were used in this study. Principal component analysis and cluster analysis were used to identify dietary patterns and microbiome community types, respectively. The association between dietary pattern and CIN risk was assessed using multivariable logistic regression analysis. The combined effect of dietary pattern and microbiome on CIN risk was determined using relative excess risk due to interaction (RERI) and synergy index (S). RESULTS: Two dietary patterns and four community types were identified: prudent diet characterized by higher intake of vegetables and fishes; semi-Western diet characterized by higher intake of bread, dairy products, eggs, and soft drinks and relatively higher fat intake ratio; and Lactobacillus crispatus-, L. iners-, Atopobium vaginae-, and Prevotella bivia-dominant types. The high-scoring group of participants with a semi-Western diet had a higher risk of CIN (odds ratio [OR] 3.44, 95% confidence interval [CI] 1.11-10.7, p = 0.03), compared with the low or medium-scoring group of those with a semi-Western diet. L. iners-dominant (OR 6.39, 95% CI 1.52-26.7, p = 0.01) and A. vaginae-dominant (OR 4.99, 95% CI 1.17-21.3, p = 0.03) dominant types had a higher risk of CIN, compared with the L. crispatus-dominant type. The synergistic effect of semi-Western diet and A. vaginae-dominant type on CIN risk was observed (OR 20.8, 95% CI 2.21-195.6, p = 0.01, RERI/S 9.64/1.96). CONCLUSIONS: Our findings suggest that semi-Western diet and its combination with A. vaginae-dominant microflora may represent an important risk factor for cervical neoplasia.

Colonization of the upper genital tract by vaginal bacterial species in nonpregnant women.

OBJECTIVE: The objective of the study was to evaluate the upper genital tract (UGT) presence of vaginal bacterial species using sensitive molecular methods capable of detecting fastidious bacterial vaginosis (BV)-associated bacteria. STUDY DESIGN: Vaginal swabs were collected prior to hysterectomy. The excised uterus was sterilely opened and swabs collected from the endometrium and upper endocervix. DNA was tested in 11 quantitative polymerase chain reaction (PCR) assays for 12 bacterial species: Lactobacillus iners, L crispatus, L jensenii, Gardnerella vaginalis, Atopobium vaginae, Megasphaera spp, Prevotella spp, Leptotrichia/Sneathia, BVAB1, BVAB2, BVAB3, and a broad-range16S ribosomal ribonucleic acid gene assay. Endometrial fluid was tested with Luminex and an enzyme-linked immunosorbent assay for cytokines and defensins and tissue for gene expression of defensins and cathelicidin. RESULTS: We enrolled 58 women: mean aged 43±7 years, mostly white (n=46; 79%) and BV negative (n=43; 74%). By species-specific quantitative PCR, 55 (95%) had UGT colonization with at least 1 species (n=52) or were positive by 16S PCR (n=3). The most common species were L iners (45% UGT, 61% vagina), Prevotella spp (33% UGT, 76% vagina) and L crispatus (33% UGT, 56% vagina). Median quantities of bacteria in the UGT were lower than vaginal levels by 2-4 log10 ribosomal ribonucleic acid gene copies per swab. There were no differences in the endometrial inflammatory markers between women with no bacteria, Lactobacillus only, or any BV-associated species in the UGT. CONCLUSION: Our data suggest that the endometrial cavity is not sterile in most women undergoing hysterectomy and that the presence of low levels of bacteria in the uterus is not associated with significant inflammation.

[Risk factors for bacterial vaginosis - a 3-year study in four medical centers].

BACKGROUND: Bacterial vaginosis (BV) is the most common infection of the lower genital tract among women of reproductive age. The purpose of this study was to determine the frequency of BV and the important etiological agent Gardnerella vaginalis in Bulgarian patients of different age groups, as well as, the risk factors forBV METHODS: One thousand five hundred and twenty-three (1523) women aged 16 to 45 years without previous antimicrobial therapy were included in this study. The methods used were: scoring of Gram staining of vaginal smear and polymerase chain reaction (PCR) assay for G. vaginalis. RESULTS: Positive for BV according to the microscopic examination criteria were 47.80% from the 1523 tested women. In 88.87% from the positive samples G. vaginalis was detected using PCR, thus demonstrating high comparability of the both methods and the leading role of G. vaginalis. The most affected with BV is the age group of 21-25 years (33.21% of all subjects with BV). The most important detected risk factors were: smoking (more than 55% with BV), single marital status (only 15% with BV are married), more than I sexual partner (more than 36% had changed the sexual partner), early onset of sexual activity (75% of B V asocciated subjects started their sexual activity by the age 15-18 years). CONCLUSIONS: The established early age for aquiring BV among Bulgarian women is very important and alarming sign. This is the first study on the etiological role of G. vaginalis and on the risk factors for BV in Bulgaria.

Characterization of the vaginal microflora in health and disease.

BACKGROUND: Bacterial vaginosis (BV) is an imbalance of the vaginal bacterial microbiota and its aetiology is still unknown. Our aims were to investigate the diagnostic potential of species/genus specific quantitative PCR (qPCR) for bacteria present in swabs and first-void urine (FVU) samples using Nugent's and Claeys' criteria and 454 sequencing of the vaginal microbiome as reference. METHODS: Self-collected swabs, vaginal smears and FVU were obtained from 177 women from Greenland (Study I and III) and physician-collected vaginal swabs and smears were obtained from 163 Swedish women (Study II). BV was diagnosed by Nugent's criteria in Study I and III and by Amsel's criteria in Study II. The vaginal swabs and FVU samples were analysed by qPCR for selected vaginal bacteria in all three studies and for four sexually transmitted infections (STIs) in Study I. RESULTS: Study I: STIs were common in women from Greenland and BV was found in 45% of these women but was not associated with individual STIs. In multivariate logistic analysis, Atopobium vaginae and Prevotella spp. were both independently associated with BV in swabs. BV could be subdivided into clusters dominated by a single or a few species together. Seven vaginal bacteria (A. vaginae, Prevotella spp. Gardnerella vaginalis, Bacterial vaginosis associated bacterium (BVAB) 2, Eggerthella-like bacterium, Leptotrichia amnionii and Megasphaera type 1) had areas under the receiver operating characteristic (ROC) curve > 85% in swabs, suggesting that they were good predictors of BV according to Nugent. Study II: For the majority of species/genera, the kappa values indicated fair to good agreement when their presence was determined by 454 pyrosequencing versus real-time PCR. The same seven vaginal bacteria as found in Study I, had areas under the ROC-curve > 85% in swabs from Swedish women, demonstrating a good diagnostic accuracy for BV according to Amsel. Study III: In a multivariate model, Megasphaera type 1 and Prevotella spp. remained significantly associated with BV in FVU samples. A linear regression analysis showed good agreement between bacterial load from swabs and FVU, but Prevotella spp. could be detected in high numbers in a few FVU samples without being present in swabs. After applying ROC curve analysis, the same seven vaginal bacteria as previously mentioned showed good prediction for BV according to Nugent in FVU. BV could be detected with comparable sensitivity in FVU and vaginal swabs. CONCLUSION: BV can be diagnosed by molecular methods performed either on swabs or urine but it is important to apply thresholds in order to improve the accuracy of the diagnosis. Furthers it was possible to identify clusters of BV dominated by single or paired bacteria, and these clusters could classify BV into subgroups, providing a more detailed understanding of the condition. Seven vaginal bacteria were highly accurate for BV diagnosis both in swabs and FVU. Finally a good agreement between Nugent and Claeys was found.